Microtiter plate assay for the measurement of glutathione and glutathione disulfide in large numbers of biological samples.
By combining the least complicated and expedient methods of sample handling with the sensitivity and specificity of the GSH assay by enzymatic recycling and the small volumes and software capabilities of microtiter plate technology we have devised a rapid, sensitive, and easy assay for GSH and GSSG in biological samples. The assay is sensitive to 5 pmol in sample volumes of 50 microliters, although other volumes could be used. The use of a computer-driven microplate with software capable of linear kinetic data storage and analysis on each well, Maxline series microplate readers and Softmax software, enables the user not only to assay large numbers of samples per day but also to have immediate calculated results. We suggest by examples that measurements of total GSH as well as changes in GSH:GSSG in vitro and in vivo are feasible with this technology.
Baker
M A
MA
Department of Radiation Oncology, University of Pennsylvania, Philadelphia 19104.
Cerniglia
G J
GJ
Zaman
A
A
D000818:Animals; D001330:Electronic Data Processing; D005978:Glutathione / Q000031:analogs & derivatives* / Q000032:analysis; D019803:Glutathione Disulfide; D005980:Glutathione Reductase / Q000032:analysis*; D006801:Humans; D007700:Kinetics; D051379:Mice; D008819:Mice, Nude; D009928:Organ Specificity; D012680:Sensitivity and Specificity; D014407:Tumor Cells, Cultured / Q000201:enzymology
D005980:Glutathione Reductase; D005978:Glutathione; D019803:Glutathione Disulfide